title: Purification and characterization of glucose-6-phosphate dehydrogenase from Pseudomonas W6.
abstract: Glucose-6-phosphate dehydrogenase (EC 1.1.1.49) has been purified from methanol grown Pseudomonas W6 by a simple procedure involving dye-ligand affinity chromatography on Cibacronblue F3G-A-Sephadex and Procion Red HE-3B-Sepharose. The purification procedure yielded a homogeneous enzyme with (1) high specific activity of 390 and 500 units/mg with NADP and NAD, respectively, and (2) low concentrations of contaminating activities. The molecular mass of the native enzyme was estimated to be 123 +/- 5 kDa. For the polypeptide chain after SDS denaturation a molecular mass of about 61 kDa was calculated. The kinetic behaviour of glucose-6-phosphate dehydrogenase exhibiting activity with either NADP or NAD was studied with respect to the substrate and coenzyme affinities and to ATP inhibition of enzyme activity. The applicability of prepared glucose-6-phosphate dehydrogenase as an auxiliary enzyme in clinical tests is discussed.
authors.affiliation: Institute of Biochemistry, Karl Marx University, Leipzig, GDR.
authors.forename: R
authors.initials: R
authors.lastname: Reuter
authors.forename: M
authors.initials: M
authors.lastname: Naumann
authors.forename: P
authors.initials: P
authors.lastname: Metz
authors.forename: G
authors.initials: G
authors.lastname: Kopperschläger
mesh_terms: D002846:Chromatography, Affinity; D003067:Coenzymes / Q000302:isolation & purification; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification*; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D011489:Protein Denaturation; D011549:Pseudomonas / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D003067:Coenzymes; D009243:NAD; D005954:Glucosephosphate Dehydrogenase